Figure 8.

GM could induce the formation of ROS, the damage of mitochondrial membrane potential, an increase of autophagy and some degree of apoptosis in auditory HCs of P4 C57BL/6 mice in vitro, which could be effectively decreased by NAC. (A) Results of Rhodamine 123 staining revealed the control HCs with dim florescence, GM treated HCs with much stronger fluorescence which could be decreased by use of NAC. The florescence intensity of Rhodamine 123 was in accordance to the DCFH-DA staining, which reflected the level of ROS in tissues. (B) GM could cause high level of LC3B (green arrows) and severe HC damages (yellow arrows), which could be decreased by NAC. (C) Immunofluorescence staining revealed that there was some activated caspase 3 in HCs after GM treatment for 24 h (blue arrows). ROS scavenger NAC could decrease the level of cleaved-caspase 3 obviously. (D) TUNEL staining showed that GM could cause some degree of TUNEL positive nuclei, which was inhibited by NAC (white arrows). (E) Western blotting analysis of LC3B-II, p53 and cleaved-caspase 3 revealed the same results with immunofluorescence. Results were shown as mean ± SEM, *p < 0.05, **p < 0.01.